Entamoeba invadens: Identification of ADF/cofilin and their expression analysis in relation to encystation and excystation

Makioka, Asao; Kumagai, Masahiro; Hiranuka, Kazushi; Kobayashi, Seiki; Takeuchi, Tsutomu

doi:10.1016/j.exppara.2010.07.018

Cited by 19 publications

(19 citation statements)

References 20 publications

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“…This suggests their involvement in the early phase of encystation. This is in contrast to ADF/cofilin because our previous study demonstrated that the expression of EiCfl-2 decreased during encystation (Makioka et al 2010). The elevated mRNA expression in the early phase of encystation is already known for E. invadens chitinases (Villagómez-Castro et al 1992;de la Vega et al 1997), and we also demonstrated the elevated mRNA level of EiChit2, which was most expressed in trophozoites among E. invadens chitinases in the early phase of encystation (Makioka et al 2011).…”

Section: Discussionmentioning

confidence: 53%

“…We previously demonstrated that excystation was enhanced by cytochalasin D cultured in growth medium but not in encystation medium (Makioka et al 2001) and further demonstrated that mRNA of EiCfl-1 and EiCfl-3 but not EiCfl-2 increased remarkably in the presence of the drug (Makioka et al 2010). Then, we studied whether the Fig.…”

Section: Mrna Expression Of Eipfn Through Encystation and Excystationmentioning

confidence: 99%

“…Previously, we found that cytochalasin D, an inhibitor of actin polymerization, unexpectedly enhanced excystation of E. invadens in vitro (Makioka et al 2001). The mechanism is still unclear, but we recently demonstrated that this enhancement by the drug is related to the remarkably increased mRNA level of two ADF/cofilins (EiCfl-1 and EiCfl-3) but not EiCfl-2 (Makioka et al 2010). Therefore, it is of interest to determine whether this enhancement by the drug is also related to the expression of E. invadens profilins.…”

Section: Introductionmentioning

confidence: 99%

“…For these reasons, E. invadens has been commonly accepted as an alternative and representative model for the study of encystation and excystation of Entamoeba. Using this in vitro system of E. invadens, we recently investigated an actin depolymerizing factor (ADF)/cofilin (Cfl), one of the important actin-binding proteins for reorganization of actin cytoskeleton, in which we identified Cfl genes and analyzed their expression in the encystation and excystation (Makioka et al 2010).…”

Section: Introductionmentioning

confidence: 99%

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Expression analysis of Entamoeba invadens profilins in encystation and excystation

et al. 2011

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Cell motility by actin cytoskeleton is essential for differentiation processes of excystation and encystation of Entamoeba. We recently studied an actin depolymerizing factor (ADF)/cofilin (Cfl) of Entamoeba invadens (Ei), and demonstrated its contribution to the encystation and excystation of E. invadens through actin cytoskeletal reorganization. Profilin is also an actin-binding protein but its function is different from that of Cfl in actin assembly. This study investigated E. invadens profilins in relation to encystation and excystation which were induced in axenic culture systems. A homology search of the E. invadens genome database and molecular cloning identified four profilins of the parasite named EiPFN1, EiPFN2, EiPFN3, and EiPFN4. There were also multiple genes of profilin in Entamoeba histolytica (Eh) and Entamoeba dispar (Ed), each of which had three profilins. A search for conserved domains revealed that these profilins of Entamoeba had actin, phosphoinositide, and poly-proline binding sites. Phylogenetic analysis revealed that EiPFN3 and EiPFN4 formed the same clades including EhPFN3 and EdPFN3, and EhPFN2 and EdPFN2, respectively, while EiPFN1 and EiPFN2 were separated from EhPFN1 and EdPFN1. Rabbit anti-EiPFN1 serum reacted with recombinant EiPFN3 and EiPFN4 but not EiPFN2, and also reacted with EiPFN in lysates of cysts and trophozoites. Immunofluorescence staining with this antiserum showed co-localization of EiPFN with actin beneath the cell membrane through the life stages and also showed cytoplasmic localization. Both proteins proved to be rich in pseudopodia of trophozoites. Real-time RT-PCR showed that the mRNA level of EiPFN1 and EiPFN4 in trophozoites was comparable but that of EiPFN2 and EiPFN3 was very low. During encystation, the mRNA expression of EiPFN1 and EiPFN4 increased remarkably in the early phase much higher than that of EiPFN2 and EiPFN3. Then, the expression of all four PFNs sharply decreased in the later phase. This was in contrast to the sharp decrease in the mRNA level of EiCfl-2 during encystation in our previous study. In cysts, EiPFN1 was most abundantly expressed and EiPFN4 was at a lower level, while the expressions of EiPFN2 and EiPFN3 were virtually absent. Following the induction of excystation, mRNA levels of EiPFN1, EiPFN2, and EiPFN4 in cysts 5 h after induction were significantly higher than those in cysts before induction, while that of EiPFN3 was slightly higher than before induction. The mRNAs of EiPFN1 increased most extensively when the excystation was induced in the presence of cytochalasin D. Small interfering RNA (siRNA) to EiPFN1 inhibited both encystation and excystation but not growth. These findings demonstrate different expression of EiPFNs and the contribution of EiPFN to the encystation and excystation.

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Section: Discussionmentioning

confidence: 53%

Section: Mrna Expression Of Eipfn Through Encystation and Excystationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Expression analysis of Entamoeba invadens profilins in encystation and excystation

et al. 2011

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“…Recently, three genes encoding proteins of the ADP/cofilin family of Entamoeba invadens have been studied. One of them was repressed during encystment, while the two others were induced during excystment (33).…”

Section: The Cytoskeletonmentioning

confidence: 99%

Cellular, Biochemical, and Molecular Changes during Encystment of Free-Living Amoebae

Fouque

Trouilhé

Thomas³

et al. 2012

Eukaryot Cell

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ABSTRACTFree-living amoebae are protozoa found in soil and water. Among them, some are pathogenic and many have been described as potential reservoirs of pathogenic bacteria. Their cell cycle is divided into at least two forms, the trophozoite and the cyst, and the differentiation process is named encystment. As cysts are more resistant to disinfection treatments than trophozoites, many studies focused on encystment, but until recently, little was known about cellular, biochemical, and molecular modifications operating during this process. Important signals and signaling pathways at play during encystment, as well as cell responses at the molecular level, have been described. This review summarizes our knowledge and focuses on new findings.

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Different structure and mRNA expression of Entamoeba invadens chitinases in the encystation and excystation

et al. 2011

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Entamoeba invadens: Identification of ADF/cofilin and their expression analysis in relation to encystation and excystation

Cited by 19 publications

References 20 publications

Expression analysis of Entamoeba invadens profilins in encystation and excystation

Expression analysis of Entamoeba invadens profilins in encystation and excystation

Cellular, Biochemical, and Molecular Changes during Encystment of Free-Living Amoebae

Different structure and mRNA expression of Entamoeba invadens chitinases in the encystation and excystation

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